High-Throughput Screening for Extracellular Inhibitors of the FLT3 Receptor Tyrosine Kinase Reveals Chemically Diverse and Druggable Negative Allosteric Modulators.

Autor: Hany R; Plate-forme de Chimie Biologique Intégrative de Strasbourg (PCBIS), UAR3286 CNRS-Université de Strasbourg, Institut du Médicament de Strasbourg, ESBS Pôle API, Bld Sébastien Brant, 67412 Illkirch Cedex, France., Leyris JP; Institut des Neurosciences de Montpellier (INM), INSERM, Institut National de la Santé et de la Recherche Médicale, UMR1051, Hôpital Saint-Eloi, 34000 Montpellier, France.; Université de Montpellier, 34000 Montpellier, France.; BIODOL Therapeutics, CAP Alpha, 34830 Clapiers, France., Bret G; Laboratoire d'Innovation Thérapeutique (LIT), UMR7200 CNRS-Université de Strasbourg, 67400 Illkirch, France., Mallié S; Institut des Neurosciences de Montpellier (INM), INSERM, Institut National de la Santé et de la Recherche Médicale, UMR1051, Hôpital Saint-Eloi, 34000 Montpellier, France.; Université de Montpellier, 34000 Montpellier, France., Sar C; Institut des Neurosciences de Montpellier (INM), INSERM, Institut National de la Santé et de la Recherche Médicale, UMR1051, Hôpital Saint-Eloi, 34000 Montpellier, France.; Université de Montpellier, 34000 Montpellier, France., Thouaye M; Institut des Neurosciences de Montpellier (INM), INSERM, Institut National de la Santé et de la Recherche Médicale, UMR1051, Hôpital Saint-Eloi, 34000 Montpellier, France.; Université de Montpellier, 34000 Montpellier, France., Hamze A; Université Paris-Saclay, CNRS, BioCIS, 92290, Châtenay-Malabry, France., Provot O; Université Paris-Saclay, CNRS, BioCIS, 92290, Châtenay-Malabry, France., Sokoloff P; BIODOL Therapeutics, CAP Alpha, 34830 Clapiers, France., Valmier J; Institut des Neurosciences de Montpellier (INM), INSERM, Institut National de la Santé et de la Recherche Médicale, UMR1051, Hôpital Saint-Eloi, 34000 Montpellier, France.; Université de Montpellier, 34000 Montpellier, France., Villa P; Plate-forme de Chimie Biologique Intégrative de Strasbourg (PCBIS), UAR3286 CNRS-Université de Strasbourg, Institut du Médicament de Strasbourg, ESBS Pôle API, Bld Sébastien Brant, 67412 Illkirch Cedex, France., Rognan D; Laboratoire d'Innovation Thérapeutique (LIT), UMR7200 CNRS-Université de Strasbourg, 67400 Illkirch, France.
Jazyk: angličtina
Zdroj: ACS chemical biology [ACS Chem Biol] 2022 Mar 18; Vol. 17 (3), pp. 709-722. Date of Electronic Publication: 2022 Feb 28.
DOI: 10.1021/acschembio.2c00048
Abstrakt: Inhibiting receptor tyrosine kinases is commonly achieved by two main strategies targeting either the intracellular kinase domain by low molecular weight compounds or the extracellular ligand-binding domain by monoclonal antibodies. Identifying small molecules able to inhibit RTKs at the extracellular level would be highly desirable to gain exquisite selectivity but is believed to be challenging owing to the size of RTK endogenous ligands (cytokines, growth factors) and the topology of RTK extracellular domains. We here report the high-throughput screening of the French Chemical Library (48K compounds) for extracellular inhibitors of the Fms-like tyrosine kinase 3 (FLT3) receptor tyrosine kinase, by a homogeneous time-resolved fluorescence competition assay. A total of 679 small molecular weight ligands (1.4%) were confirmed to strongly inhibit (>75%) the binding of the fluorescent labeled FLT3 ligand (FL cytokine) to FLT3 overexpressed in HEK-293 cells, at two different concentrations (5 and 20 μM). Concentration-response curves, obtained for 111 lead-like molecules, confirmed the unexpected tolerance of the FLT3 extracellular domain for low molecular weight druggable inhibitors exhibiting submicromolar potencies, chemical diversity, and promising pharmacokinetic properties. Further investigation of one hit confirmed inhibitory properties in dorsal root ganglia neurons and in a mouse model of neuropathic pain.
Databáze: MEDLINE
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